吉西他滨对人骨肉瘤细胞增殖及肿瘤转移相关基因1 mRNA表达的影响

Effect of Gemcitabine on the Proliferation and MTA1 mRNA Expression in Human Osteosarcoma MG- 63 Cells

目的探讨吉西他滨对体外培养的人骨肉瘤细胞增殖的影响及其可能的作用机制。方法体外传代培养人骨肉瘤MG-63细胞,实验时取对数生长期的细胞。将不同浓度(0.025、0.050、0.100、0.200、0.400 mg/L)的吉西他滨分别作用于MG-63细胞24 h,用四甲基偶氮唑盐(MTT)法检测其对MG-63细胞的生长抑制率。用实时定量聚合酶链反应(PCR)检测吉西他滨作用24 h后人骨肉瘤MG-63细胞肿瘤转移相关基因1(MTA1)mRNA的相对表达量。结果 0.025、0.050、0.100、0.200、0.400 mg/L吉西他滨作用24 h后,MG-63细胞抑制率分别为(3.3±1.2)%、(3.7±1.0)%、(6.1±0.1)%、(8.0±1.9)%和(11.5±1.4)%,差异有统计学意义(F=30.568,P=0.0169);其中0.025、0.050 mg/L组MG-63抑制率均低于其他3组,0.100、0.200 mg/L组亦低于0.400 mg/L组(P<0.05)。0.100、0.200、0.400 mg/L吉西他滨作用于人骨肉瘤MG-63细胞24 h后,MTA1 mRNA相对表达量〔分别为(0.86±0.16)、(0.68±0.12)、(0.46±0.08)〕均低于空白对照组(1.01±0.15),且3个剂量吉西他滨组MTA1mRNA相对表达量依次降低,差异有统计学意义(P<0.01)。结论吉西他滨能抑制人骨肉瘤MG-63细胞增殖,且有剂量依赖性;其作用机制可能是通过下调MTA1 mRNA的表达,从而抑制MG-63细胞转移,发挥抗骨肉瘤作用。

Objective To investigate the influence of gemcitabine on the proliferation of human osteosarcoma MG -63 ceUs in vitro, and explore the mechanism. Methods Human osteosarcoma MG -63 cells were cultured serially, and the ceils at exponential phase of growth were adopted in experiment. Mono - nuclear cell direct cytotoxicity MTT assay was employed to e- valuate the inhibitory effect of gemcitabine （0. 025, 0. 050, 0. 100, 0. 200, 0. 400 nag/L） on growth of cultured human osteo- sarcoma MG - 63 cells. The mRNA expression of MTA1 after 24 - hour gemcitabine treatment was measured by real - time RT - PCR. Results After 24-hour treatment with of 0. 025, 0. 050, 0. 100, 0. 200, 0. 400 mg/L of gemcitabine, the inhibition rates of MG-63 were （3.3±1.2）%, （3.7±1.0）%, （6.1±0.1）%, （8.0±1.9）%, and （11.5±1.4）%, respective- ly. Gemcitabine significantly inhibited the proliferation of the MG - 63 cells in a dose - dependent manner （ F = 30. 568, P = 0. 0169）. The inhibition rates in 0. 025 mg/L group and 0. 050 mg/L group were significantly lower than those in the other three groups, and the inhibition rates in 0. 100 mg/L group and 0. 200 mg/L group were significantly lower than in 0. 400 mg/L group （P 〈0. 05）. After 24 - hour gemcitabine treatment, the mRNA expressions of MTA1 in 0. 100 rag/L, 0. 200 mg/L and 0. 400 mg/L groups were （0.86±0.16）, （0.68±0.12）, and （0.46±0.08）, respectively, all of which were significantly lower than that in the control group （1.01 ±0. 15）, and the differences among each gemcitabine groups were also statistically signifi-cant （ P 〈 0. 01 ）. Conclusion Gemcitabine of certain concentration has antitumor effect on osteosarcoma in vitro through inhib- iting cell proliferation, which might be related to the regulation of acetyl level of histone and the down - regulated mRNA expres- sion of MTA1. Gemeitabine might be a promising new antitumor drug to treat osteosarcoma.